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目的 :采用目前治疗绒癌最常用的药物诱导建立绒癌的耐药细胞系 ,比较不同药物引起耐药机制的差异。方法 :采用浓度梯度递增法和间歇诱导法 ,分别用 5 FU ,MTX ,VP 16诱导绒癌细胞系JEG 3,建立相应耐药细胞系 ,采用MTT法测定耐药细胞系对5 FU、MTX、KSM、VP 16和Taxol的耐药指数 ,用RT PCR测定亲本细胞及各耐药细胞系耐药相关基因如肺耐药蛋白 (LRP)、多药耐药相关蛋白 (MRP)、谷胱甘肽转移酶 (GST π)、二氢叶酸还原酶 (DHFR)和多药耐药基因 (MDR 1)的mRNA表达 ,比较各种细胞生长曲线 ,细胞群体倍增时间。结果 :JEG 3细胞系在诱导耐药细胞前即有LRP、MRP、GST π、DHFR的共表达 ,诱导耐药后上述各基因的表达无明显变化。诱导耐药前JEG 3细胞无MDR1的表达 ,用VP 16、5 FU间歇诱导的方法诱导形成的耐药细胞系有MDR 1表达 ,且耐药指数增加。结论 :JEG 3耐药细胞系耐药性的产生与MRP、LRP、GST π、DHFR表达的关系不密切 ,而与MDR1的表达有关。
Objective: To use the most commonly used drugs for the treatment of choriocarcinoma to induce the establishment of a resistant cell line of choriocarcinoma, and to compare the differences in drug resistance mechanisms caused by different drugs. METHODS: Using a concentration gradient and intermittent induction method, 5 FU, MTX, and VP 16-induced JEV 3 cell lines were established to establish drug-resistant cell lines. MTT assay was used to determine drug-resistant cell lines against 5 FU and MTX. The resistance index of KSM, VP 16 and Taxol was determined by RT PCR to detect the resistance-associated genes such as lung resistance protein (LRP), multidrug resistance-associated protein (MRP) and glutathione in parental cells and resistant cell lines. The mRNA expression of transferase (GST π), dihydrofolate reductase (DHFR) and multidrug resistance gene (MDR 1) was compared with various cell growth curves and cell population doubling time. Results: The expression of LRP, MRP, GST π and DHFR was observed in JEG 3 cell lines before induction of drug-resistant cells. No significant changes in the expression of these genes were observed after induction of drug resistance. The MDR1 expression was not found in JEG 3 cells before induction of drug resistance. MDR 1 expression was induced in the resistant cell lines induced by intermittent induction of VP 16 and 5 FU, and the resistance index increased. Conclusion : The development of drug resistance in JEG 3 cell lines is not closely related to the expression of MRP, LRP, GST π and DHFR, but related to the expression of MDR1.